Extensive regulation of enzyme activity by phosphorylation in Escherichia coli.
Extensive regulation of enzyme activity by phosphorylation in Escherichia coli.
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大肠杆菌中磷酸化对酶活性的广泛调节。
DOI：
10.1038/s41467-021-25988-4
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发表时间：
2021-09-24
影响因子：
16.6
通讯作者：
Sauer U
中科院分区：
文献类型：
--
作者：
Schastnaya E;Raguz Nakic Z;Gruber CH;Doubleday PF;Krishnan A;Johns NI;Park J;Wang HH;Sauer U
蛋白质丝氨酸/苏氨酸/酪氨酸（S/T/Y）磷酸化是真核生物中必需且频繁的翻译后修饰，但历史上被认为在细菌中不太普遍，因为发现较少的蛋白质被磷酸化，并且大多数蛋白质被修饰至较低程度。最近的蛋白质组学研究大大扩大了磷酸化蛋白质组的大肠杆菌超过2000磷酸化位点（磷酸化位点），但提出的行动机制，只有6个磷酸化位点和健身效果被描述为38磷酸化扰动后。通过系统研究E.根据生长表型和细胞内代谢组谱，我们发现52个突变磷酸酶中有44个具有功能。通过有效地加倍已知的功能磷酸化位点的数量，我们提供的证据表明，蛋白质磷酸化是细菌代谢的主要调节过程。结合在体外和体内实验中，我们展示了如何单磷酸调节酶的活性和调节代谢通量糖酵解，甲基乙二醛旁路，乙酸代谢和戊糖磷酸和恩特纳-杜道夫途径之间的分裂，通过机制，包括屏蔽底物结合位点，限制结构动力学，并破坏相关的活性在体内的相互作用。磷酸化是真核生物中一种重要的翻译后修饰，而原核生物的磷酸化蛋白质组研究则是最近才开始的。在这里，Schastnaya等人突变了23个E.大肠杆菌酶和应用代谢组学提供证据的功能相关性的细菌磷酸化事件。
Protein serine/threonine/tyrosine (S/T/Y) phosphorylation is an essential and frequent post-translational modification in eukaryotes, but historically has been considered less prevalent in bacteria because fewer proteins were found to be phosphorylated and most proteins were modified to a lower degree. Recent proteomics studies greatly expanded the phosphoproteome of Escherichia coli to more than 2000 phosphorylation sites (phosphosites), yet mechanisms of action were proposed for only six phosphosites and fitness effects were described for 38 phosphosites upon perturbation. By systematically characterizing functional relevance of S/T/Y phosphorylation in E. coli metabolism, we found 44 of the 52 mutated phosphosites to be functional based on growth phenotypes and intracellular metabolome profiles. By effectively doubling the number of known functional phosphosites, we provide evidence that protein phosphorylation is a major regulation process in bacterial metabolism. Combining in vitro and in vivo experiments, we demonstrate how single phosphosites modulate enzymatic activity and regulate metabolic fluxes in glycolysis, methylglyoxal bypass, acetate metabolism and the split between pentose phosphate and Entner-Doudoroff pathways through mechanisms that include shielding the substrate binding site, limiting structural dynamics, and disrupting interactions relevant for activity in vivo. While phosphorylation is an essential post-translational modification in eukaryotes only recently the phosphoproteome of prokaryotes has been provided. Here, Schastnaya et al. mutate 52 phosphosites on 23 E. coli enzymes and apply metabolomics to provide evidence for the functional relevance of bacterial phosphorylation events.
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影响因子：
--
作者：
Burnell JN
通讯作者：
Burnell JN
DOI：
10.1073/pnas.1811971115
发表时间：
2018-10-23
影响因子：
11.1
作者：
Brunk, Elizabeth;Chang, Roger L.;Lewis, Nathan E.
通讯作者：
Lewis, Nathan E.
影响因子：
7.4
作者：
Fuhrer, Tobias;Heer, Dominik;Zamboni, Nicola
通讯作者：
Zamboni, Nicola
影响因子：
3.7
作者：
Lacour, Soline;Bechet, Emmanuelle;Grangeasse, Christophe
通讯作者：
Grangeasse, Christophe
影响因子：
2.6
作者：
Lacour, Soline;Doublet, Patricia;Grangeasse, Christophe
通讯作者：
Grangeasse, Christophe